Title
Genetically encoded red fluorescent copper(I) sensors for cellular copper(I) imaging
Author
Junyi Liang, Lele Guo, Yin Ding, Lei Xia, Yan Shen, Meng Qin, Qiang Xu, Yi Cao, Wei Wang
Year
2014
Journal
Biochemical and Biophysical Research Communications
Abstract
Copper ranks among the most important metal ions in living organism, owing to its key catalytic effect in a range of biochemical processes. Dysregulation of in vivo copper(I) metabolism is extremely toxic and would cause serious diseases in human, such as Wilson’s and Menkes. Thus, it would be highly valuable to have a proper approach to monitor the dynamics of copper(I) in vivo, as it is directly related to the onset of human copper(I)-related diseases. Under these circumstance, developing fluorescent protein based copper(I) sensors is highly demanded. However, these established sensors are mostly based on green or yellow FPs. Fluorescent copper(I) sensors with a spectra in the red range are more desirable due to lower phototoxicity, less auto-fluorescent noise and better penetration of red light. In the present work, we grafted a special red FP into three different location of a copper(I) binding protein, and generate a series of red fluorescent copper(I) sensors. Despite their limited in vivo sensitivity toward copper(I), these sensors are viable for cellular copper(I) imaging. Furthermore, these red fluorescent copper(I) sensors are a good starting point to develop superior copper(I) biosensors capable of imaging copper(I) fluctuations within a truly biologically relevant concentration, and further effort to realize this endeavor is under way.
Instrument
FP-6500
Keywords
Fluorescence, Protein structure, Ligand binding, Sensors, Biochemistry, Materials